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Improvements in slide preparation from archival material for automated DNA measurement by image analysis
Objective:
To optimize slide preparation for DNA content measurement by automated image analysis and to obtain a rapid and easy method for routine use.
Study Design:
Some improvements in previously described methods were achieved: automatic dewaxing, reduction of washing times, accurate adjustment of final concentration of nuclei, sedimentation and temperate drying of nuclei.
Results:
The complete preparation of 12 slides required four intermittent working hours. Accurate adjustment of the final concentration and sedimentation of nuclei resulted in a constant and homogeneous distribution of nuclei. Cell loss was prevented and the most fragile structures preserved.
Conclusion:
High-quality preparations of nuclei lead to high-resolution histograms obtained from a large collection of events. This promotes automated image analysis method instead of flow cytometry when only archival material is available for DNA content measurement and proliferating fraction evaluation.